Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Protocol
  • Published:

Silver staining DNA in polyacrylamide gels

Abstract

This protocol describes a simple silver staining method used to visualize DNA fragments and other organic molecules with unsurpassed detail following traditional polyacrylamide gel electrophoresis (PAGE). Sensitivity rivals radioisotopic methods and DNA in the picogram range can be reliably detected. The described protocol is fast (1 h) and is implemented using readily available chemicals and materials. To achieve the sensitivity and visual clarity expected, quality reagents and clean handling are important. The updated protocol described here is based on the widely used method of Bassam et al. (1991), but provides improved image contrast and less risk of staining artefacts.

This is a preview of subscription content, access via your institution

Access options

Buy this article

Prices may be subject to local taxes which are calculated during checkout

Figure 1: I DNA silver staining of polyacrylamide gel electrophoresis separated arbitrary primed DNA amplification products: The figure shows each of the six treatments in order (af), as described in Table 2.

Similar content being viewed by others

References

  1. Switzer, R.C. III, Merril, C.R. & Shifrin, S. A highly sensitive silver stain for detecting proteins and peptides in polyacrylamide gels. Anal. Biochem. 98, 231–237 (1979).

    Article  CAS  PubMed  Google Scholar 

  2. Merril, C.R., Dunau, M.L. & Goldman, D. A rapid sensitive silver stain for polypeptides in polyacrylamide gels. Anal. Biochem. 110, 201–207 (1981).

    Article  CAS  PubMed  Google Scholar 

  3. Heukeshoven, J. & Dernick, R. Simplified method for staining of proteins in polyacrylamide gels and the mechanism of silver staining. Electrophoresis 6, 103–112 (1985).

    Article  CAS  Google Scholar 

  4. Somerville, L.L. & Wang, K. The ultrasensitive silver 'protein' stain also detects nanograms of nucleic acids. Biochem. Biophys. Res. Commun. 102, 53–58 (1981).

    Article  CAS  PubMed  Google Scholar 

  5. Boulikas, T. & Hancock, R. A highly sensitive technique for staining DNA and RNA in polyacrylamide gels using silver. J. Biochem. Biophys. Methods 4, 219–228 (1981).

    Article  Google Scholar 

  6. Tsai, C.M. & Frasch, C.E. A sensitive silver stain for detecting lipopolysaccharides in polyacrylamide gels. Anal. Biochem. 119, 115–119 (1982).

    Article  CAS  PubMed  Google Scholar 

  7. Dubray, G. & Bezard, G. A highly sensitive periodic acid-silver stain for 1,2-diol groups of glycoproteins and polysaccharides in polyacrylamide gels. Anal. Biochem. 119, 325–329 (1982).

    Article  CAS  PubMed  Google Scholar 

  8. Caetano-Anollés, G., Bassam, B.J. & Gresshoff, P.M. DNA amplification fingerprinting using very short arbitrary oligonucleotide primers. BioTechnology (NY) 9, 553–557 (1991).

    Google Scholar 

  9. Bassam, B.J., Caetano-Anollés, G. & Gresshoff, P.M. Fast and sensitive silver staining of DNA in polyacrylamide gels. Anal. Biochem. 196, 80–83 (1991).

    Article  CAS  PubMed  Google Scholar 

  10. Bassam, B.J. & Bentley, S. Electrophoresis of polyester-backed polyacrylamide gels. Biotechniques 19, 568–573 (1995).

    CAS  PubMed  Google Scholar 

  11. Merril, C.R. Silver staining of proteins and DNA. Nature 343, 779–780 (1990).

    Article  CAS  PubMed  Google Scholar 

  12. Rabilloud, T. Mechanisms of protein silver staining in polyacrylamide gels: a 10-year synthesis. Electrophoresis 10, 785–794 (1990).

    Article  Google Scholar 

  13. Guillemette, J.G. & Lewis, P.N. Detection of subnanogram quantities of DNA and RNA on native and denaturing polyacrylamide and agarose gels by silver staining. Electrophoresis 4, 92–94 (1983).

    Article  CAS  Google Scholar 

  14. Kolodny, G.M. An improved method for increasing the resolution and sensitivity of silver staining of nucleic acid bands in polyacrylamide gels. Anal. Biochem. 138, 66–67 (1984).

    Article  CAS  PubMed  Google Scholar 

  15. Beidler, J.L., Hilliard, P.R. & Rill, R.L. Ultrasensitive staining of nucleic acids with silver. Anal. Biochem. 126, 374–380 (1982).

    Article  CAS  PubMed  Google Scholar 

  16. Goldman, D. & Merril, C.R. Silver staining of DNA in polyacrylamide gels: linearity and effect of fragment size. Electrophoresis 3, 24–26 (1982).

    Article  CAS  Google Scholar 

  17. Merril, C.R., Harrington, M. & Alley, V. A photodevelopment silver stain for the rapid visualization of proteins separated on polyacrylamide gels. Electrophoresis 5, 289–297 (1984).

    Article  CAS  Google Scholar 

  18. Blum, H., Beier, H. & Gross, H.J. Improved silver staining of plant proteins, RNA and DNA in polyacrylamide gels. Electrophoresis 8, 93–99 (1987).

    Article  CAS  Google Scholar 

  19. Merril, C.R., Goldman, D., Sedman, S.A. & Ebert, M.H. Ultrasensitive stain for proteins in polyacrylamide gels shows regional variation in cerebrospinal fluid proteins. Science 211, 1437–1438 (1981).

    Article  CAS  PubMed  Google Scholar 

  20. Kruchinina, N.G. & Gresshoff, P.M. Detergent affects silver sequencing. Biotechniques 17, 280–282 (1994).

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank the Australian Research Council Centre of Excellence grant scheme (CEO348212) and associated support for the Queensland State Government, the UQ Strategic Fund and present and past members of the laboratory. Ms. Johanna Hadler and Dr. Paul Scott (both CILR Brisbane) are thanked for beta-testing the new procedure.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Peter M Gresshoff.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Bassam, B., Gresshoff, P. Silver staining DNA in polyacrylamide gels. Nat Protoc 2, 2649–2654 (2007). https://doi.org/10.1038/nprot.2007.330

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nprot.2007.330

This article is cited by

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing